| Objective To observe the effect of novel cannabinoid type 1 receptor antagonist ZH-101-S on renal mitochondria, expression of VDAC, Mfn-2 and Cyto C in kidney of rats with diabetic nephropathy, and to investigate the protective effect and mechanism of ZH-101-S on the kidney of diabetic nephropathy rats.Methods Diabetic rats model was induced by intraperitoneal injection of streptozotocin.Models were randomly and equally divided into four groups following with non-treated diabetic rats model( group DM), diabetic rats with different concentrations of CB1 receptor antagonist group:diabetic rats plus ZH-101-S0.3mg/kg(low dose group), diabetic rats plus ZH-101-S 1.0mg/kg(middle dose group)and diabetic rats plus ZH-101-S 3.0mg/kg(high dose group). The normal rats as normal control group(group NC).After 8 weeks, the excretion of serum creatinine(Scr), serum urea nitrogen(BUN), urinary albumin excretion rate(UAER) were detected. The level of malondialdehyde( MDA) and the activity of superoxide dismutase(SOD) in renal tissue were detected. The pathological changes of renal tissue were observed by HE staining. The morphological changes of mitochondria were observed by electron microscope.Additionally,renal mitochondrial swelling degree was measured among the different groups. The expressions of voltage-dependent anion channel(VDAC), mitofusin-2 and cytochrome C(Cyto C) protein in renal tissue of rats were examined by Western blot.Results1. Compared with group NC, in group DM, the UAER, Scr and BUN were significantly increased(P<0.01 or P<0.05), Compared with the group DM, in high dose group, the UAER, Scr and BUN were significantly decreased( P<0.01 or P<0.05).2. Compared with group NC, in group DM, the SOD activity in renal tissue was decreased(P<0.01), and the level of MDA was increased(P<0.05). Compared with group DM, in high dose group, the SOD activity in renal tissue was increased(P<0.01), and the level of MDA was decreased(P<0.05).3. HE staining showed increased glomerular volume, renal tubular epithelial cell edema and mesangial matrix increased in the DM group.In DM group,the mitochondrial swelling and the diminution of cristae were observed by electron microscopy. The morphology of the intervention group was improved,in high dose group were obviously ameliorated.4. Compared with group NC, in group DM, mitochondrial swelling tendency weakened. Compared with group DM, the swelling tendency of the intervention group increased gradually with the increase of the concentration.5. Compared with group NC, in group DM, the expressions of VDAC protein was significantly decreased(P<0.01). Compared with group DM, in high dose group, the expressions of VDAC protein was significantly increased(P<0.05).6. Compared with group NC, in group DM, the expressions of Mfn-2 protein was significantly decreased(P<0.01). Compared with group DM, in high dose group, the expressions of Mfn-2 protein was significantly increased(P<0.05).7. Compared with group NC, in group DM, the expressions of Cyto C protein was significantly increased(P<0.01). Compared with group DM, in high dose group,the expressions of Cyto C protein was significantly decreased(P<0.05).Conclusion Our findings indicated that increased oxidative stress happened in the kidney of diabetic rats, mitochondrial dysfunction, the expressions of VDAC and Mfn-2 protein was significantly decreased, and the expressions of Cyto C protein was significantly increased. After plus ZH-101-S, the condition was reversed. ZH-101-S can prevent the oxidative stress, protect mitochondrial function of kidney. |
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